Certain chemical entities, compositions and methods

ABSTRACT

Provided are certain chemical entities, and methods of use to modulate skeletal myosin, skeletal actin, skeletal tropomyosin, skeletal troponin C, skeletal troponin I, skeletal troponin T, and skeletal muscle, including fragments and isoforms thereof, as well as the skeletal sarcomere, and methods of use in the treatment of obesity, sarcopenia, wasting syndrome, frailty, cachexia, muscle spasm, post-surgical and post-traumatic muscle weakness, neuromuscular disease, and other indications.

This application claims the benefit of U.S. Patent Application No.61/026,076, filed Feb. 4, 2008, which is hereby incorporated byreference.

Provided are certain chemical entities that modulate skeletal myosin,skeletal actin, skeletal tropomyosin, skeletal troponin C, skeletaltroponin I, skeletal troponin T, and skeletal muscle, includingfragments and isoforms thereof, as well as the skeletal sarcomere. Alsoprovided are certain chemical entities, pharmaceutical compositions andmethods of treatment of one or more of obesity, sarcopenia, wastingsyndrome, frailty, cachexia, muscle spasm, post-surgical andpost-traumatic muscle weakness, and neuromuscular disease.

The cytoskeleton of skeletal and cardiac muscle cells is unique comparedto that of all other cells. It consists of a nearly crystalline array ofclosely packed cytoskeletal proteins called the sarcomere. The sarcomereis elegantly organized as an interdigitating array of thin and thickfilaments. The thick filaments are composed of myosin, the motor proteinresponsible for transducing the chemical energy of ATP hydrolysis intoforce and directed movement. The thin filaments are composed of actinmonomers arranged in a helical array. There are four regulatory proteinsbound to the actin filaments, which allows the contraction to bemodulated by calcium ions. An influx of intracellular calcium initiatesmuscle contraction; thick and thin filaments slide past each otherdriven by repetitive interactions of the myosin motor domains with thethin actin filaments.

Myosin is the most extensively studied of all the motor proteins. Of thethirteen distinct classes of myosin in human cells, the myosin-II classis responsible for contraction of skeletal, cardiac, and smooth muscle.This class of myosin is significantly different in amino acidcomposition and in overall structure from myosin in the other twelvedistinct classes. Myosin-II consists of two globular head domains linkedtogether by a long alpha-helical coiled-coiled tail that assembles withother myosin-IIs to form the core of the sarcomere's thick filament. Theglobular heads have a catalytic domain where the actin binding and ATPfunctions of myosin take place. Once bound to an actin filament, therelease of phosphate (cf. ATP to ADP) leads to a change in structuralconformation of the catalytic domain that in turn alters the orientationof the light-chain binding lever arm domain that extends from theglobular head; this movement is termed the powerstroke. This change inorientation of the myosin head in relationship to actin causes the thickfilament of which it is a part to move with respect to the thin actinfilament to which it is bound. Un-binding of the globular head from theactin filament (also Ca²⁺ modulated) coupled with return of thecatalytic domain and light chain to their startingconformation/orientation completes the contraction and relaxation cycle,responsible for intracellular movement and muscle contraction.

Tropomyosin and troponin mediate the calcium effect on the interactionon actin and myosin. The skeletal troponin complex regulates the actionof several actin units at once, and is comprised of three polypeptidechains: skeletal troponin C, which binds calcium ions; troponin I, whichbinds to actin; and troponin T, which binds to tropomyosin.

Accordingly, there is a need for the development of new compounds thatmodulate skeletal muscle. There remains a need for agents that exploitnew mechanisms of action and which may have better outcomes in terms ofrelief of symptoms, safety, and patient mortality, both short-term andlong-term and an improved therapeutic index.

Provided is at least one chemical entity chosen from compounds ofFormula I:

and pharmaceutically acceptable salts and tautomers thereof, wherein

R³ is selected from optionally substituted alkyl, optionally substitutedcycloalkyl, optionally substituted aryl, optionally substitutedheteroaryl, and optionally substituted heterocycloalkyl;

R⁵ is selected from halo, hydroxy, optionally substituted alkyl,optionally substituted amino, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted aryl, optionally substitutedcycloalkyl, optionally substituted heteroaryl, optionally substitutedheterocycloalkyl, optionally substituted alkoxy, sulfonyl, sulfanyl, andsulfinyl;

R⁶ is selected from hydrogen, halo, hydroxy, lower alkyl, and lowerhaloalkyl; and

R⁷ is selected from hydrogen, optionally substituted acyl, optionallysubstituted alkyl, optionally substituted aryl, optionally substitutedcycloalkyl, optionally substituted heteroaryl, optionally substitutedaminocarbonyl, sulfonyl, sulfanyl, sulfinyl, carboxy, optionallysubstituted alkoxycarbonyl, hydroxy, and cyano.

Also provided is a pharmaceutically acceptable composition comprising apharmaceutically acceptable carrier and at least one chemical entitydescribed herein.

Also provided are methods for treating a patient having a disease chosenfrom obesity, sarcopenia, wasting syndrome, frailty, cachexia, musclespasm, post-surgical and post-traumatic muscle weakness, andneuromuscular disease, comprising administering to the patient atherapeutically effective amount of at least one chemical entitydescribed herein.

Also provided is a method of treating one or more of obesity,sarcopenia, wasting syndrome, frailty, cachexia, muscle spasm,post-surgical and post-traumatic muscle weakness, neuromuscular disease,and other indications in a mammal which method comprises administeringto a mammal in need thereof a therapeutically effective amount of atleast one chemical entity described herein or a pharmaceuticalcomposition comprising a pharmaceutically acceptable excipient, carrieror adjuvant and at least one chemical entity described herein.

Also provided is a method for treating a patient having a diseaseresponsive to modulation of one or more of skeletal myosin, skeletalactin, skeletal tropomyosin, skeletal troponin C, skeletal troponin I,skeletal troponin T, and skeletal muscle, including fragments andisoforms thereof, as well as the skeletal sarcomere in a mammal whichmethod comprises administering to a mammal in need thereof atherapeutically effective amount of at least one chemical entitydescribed herein or a pharmaceutical composition comprising apharmaceutically acceptable excipient, carrier or adjuvant and at leastone chemical entity described herein.

Also provided is a method for treating a patient having a diseaseresponsive to sensitization of one or more of skeletal myosin, skeletalactin, skeletal tropomyosin, skeletal troponin C, skeletal troponin I,skeletal troponin T, and skeletal muscle, including fragments andisoforms thereof, as well as the skeletal sarcomere in a mammal, to Ca²⁺concentration through increasing the force of contraction at low Ca²⁺relative to untreated patients, which method comprises administering toa mammal in need thereof a therapeutically effective amount of at leastone chemical entity described herein or a pharmaceutical compositioncomprising a pharmaceutically acceptable excipient, carrier or adjuvantand at least one chemical entity described herein.

Also provided is a method for treating a patient having a diseaseresponsive to inhibition of one or more of skeletal myosin, skeletalactin, skeletal tropomyosin, skeletal troponin C, skeletal troponin I,skeletal troponin T, and skeletal muscle, including fragments andisoforms thereof, as well as the skeletal sarcomere in a mammal whichmethod comprises administering to a mammal in need thereof atherapeutically effective amount of at least one chemical entitydescribed herein or a pharmaceutical composition comprising apharmaceutically acceptable excipient, carrier or adjuvant and at leastone chemical entity described herein.

Other aspects and embodiments will be apparent to those skilled in theart from the following detailed description.

As used in the present specification, the following words and phrasesare generally intended to have the meanings as set forth below, exceptto the extent that the context in which they are used indicatesotherwise.

The following abbreviations and terms have the indicated meaningsthroughout:

Ac = acetyl ADP = adenosine triphosphate ATP = adenosine triphosphateBME = beta-mercaptoethanol c- = cyclo CDI = carbonyldiimidazole DMSO =dimethyl sulfoxide (DPPF)PdCl₂ = [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) DTT = Dithiothreitol EDTA =ethylenediaminetetraacetic acid Et = ethyl EtOAc = ethyl acetate EtOH =ethanol g = gram h or hr = hour i- = iso kg or Kg = kilogram l or L =liter m/z = mass-to-charge ratio Me = methyl mg = milligram min = minutemL or ml = milliliter mmol = millimole MMP = matrix metalloproteinase MW= microwave n- = normal Ph = phenyl rpm = revolutions per minute rt orRT = room temperature s- = sec- = secondary t- = tert- = tertiary THF =tetrahydrofuran vol = volume equivalent in mL/g or L/Kg for the limitingreagent unless otherwise indicated

By “optional” or “optionally” is meant that the subsequently describedevent or circumstance may or may not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “optionally substituted alkyl”encompasses both “alkyl” and “substituted alkyl” as defined below. Itwill be understood by those skilled in the art, with respect to anygroup containing one or more substituents, that such groups are notintended to introduce any substitution or substitution patterns that aresterically impractical, synthetically non-feasible, and/or inherentlyunstable.

“Alkyl” encompasses straight chain and branched chain having theindicated number of carbon atoms, usually from 1 to 20 carbon atoms, forexample 1 to 8 carbon atoms, such as 1 to 6 carbon atoms. For exampleC₁-C₆ alkyl encompasses both straight and branched chain alkyl of from 1to 6 carbon atoms. When an alkyl residue having a specific number ofcarbons is named, all branched and straight chain versions having thatnumber of carbons are intended to be encompassed; thus, for example,“butyl” is meant to include n-butyl, sec-butyl, isobutyl and t-butyl;“propyl” includes n-propyl and isopropyl. “Lower alkyl” refers to alkylgroups having one to seven carbons. In certain embodiments, “loweralkyl” refers to alkyl groups having one to six carbons. Examples ofalkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl,sec-butyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl,2-hexyl, 3-hexyl, 3-methylpentyl, and the like. Alkylene is a subset ofalkyl, referring to the same residues as alkyl, but having two points ofattachment. Alkylene groups will usually have from 2 to 20 carbon atoms,for example 2 to 8 carbon atoms, such as from 2 to 6 carbon atoms. Forexample, C₀ alkylene indicates a covalent bond and C₁ alkylene is amethylene group.

“Alkenyl” refers to an unsaturated branched or straight-chain alkylgroup having at least one carbon-carbon double bond derived by theremoval of one molecule of hydrogen from adjacent carbon atoms of theparent alkyl. The group may be in either the cis or trans configurationabout the double bond(s). Typical alkenyl groups include, but are notlimited to, ethenyl; propenyls such as prop-1-en-1-yl, prop-1-en-2-yl,prop-2-en-1-yl (allyl), prop-2-en-2-yl; butenyls such as but-1-en-1-yl,but-1-en-2-yl, 2-methyl-prop-1-en-1-yl, but-2-en-1-yl, but-2-en-1-yl,but-2-en-2-yl, buta-1,3-dien-1-yl, buta-1,3-dien-2-yl; and the like. Incertain embodiments, an alkenyl group has from 2 to 20 carbon atoms andin other embodiments, from 2 to 6 carbon atoms. “Lower alkenyl” refersto alkenyl groups having two to six carbons.

“Alkynyl” refers to an unsaturated branched or straight-chain alkylgroup having at least one carbon-carbon triple bond derived by theremoval of two molecules of hydrogen from adjacent carbon atoms of theparent alkyl. Typical alkynyl groups include, but are not limited to,ethynyl; propynyls such as prop-1-yn-1-yl, prop-2-yn-1-yl; butynyls suchas but-1-yn-1-yl, but-1-yn-3-yl, but-3-yn-1-yl; and the like. In certainembodiments, an alkynyl group has from 2 to 20 carbon atoms and in otherembodiments, from 3 to 6 carbon atoms. “Lower alkynyl” refers to alkynylgroups having two to six carbons.

“Cycloalkyl” indicates a non-aromatic carbocyclic ring, usually havingfrom 3 to 7 ring carbon atoms. The ring may be saturated or have one ormore carbon-carbon double bonds. Examples of cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, andcyclohexenyl, as well as bridged and caged ring groups such asnorbornane.

The term “alkoxy” refers to the group —O-alkyl, including from 1 to 8carbon atoms of a straight, branched, cyclic configuration andcombinations thereof attached to the parent structure through an oxygen.Examples include methoxy, ethoxy, propoxy, isopropoxy, cyclopropyloxy,cyclohexyloxy and the like. “Lower alkoxy” refers to alkoxy groupscontaining one to six carbons.

The term “substituted alkoxy” refers to alkoxy wherein the alkylconstituent is substituted (i.e., —O-(substituted alkyl)) wherein“substituted alkyl” refers to alkyl wherein one or more (such as up to5, for example, up to 3) hydrogen atoms are replaced by a substituentindependently chosen from:

—R^(a), —OR^(b), optionally substituted amino (including —NR^(c)COR^(b),—NR^(c)CO₂R^(a), —NR^(c)CONR^(b)R^(c), —NR^(b)C(NR^(c))NR^(b)R^(c),—NR^(b)C(NCN)NR^(b)R^(c), and —NR^(c)SO₂R^(a)), halo, cyano, nitro, oxo(as a substitutent for cycloalkyl, heterocycloalkyl, and heteroaryl),optionally substituted acyl (such as —COR^(b)), optionally substitutedalkoxycarbonyl (such as —CO₂R^(b)), aminocarbonyl (such as—CONR^(b)R^(c)), —OCOR^(b), —OCO₂R^(a), —OCONR^(b)R^(c), OCONR^(b)R^(c),—OP(O)(OR^(b))OR^(c), sulfanyl (such as SR^(b)), sulfinyl (such as—SOR^(a)), and sulfonyl (such as —SO₂R^(a) and —SO₂NR^(b)R^(c)),

where R^(a) is chosen from optionally substituted C₁-C₆ alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, and optionally substituted heteroaryl;

R^(b) is chosen from H, optionally substituted C₁-C₆ alkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted aryl, and optionally substituted heteroaryl; and

R^(c) is independently chosen from hydrogen and optionally substitutedC₁-C₄ alkyl; or

R^(b) and R^(c), and the nitrogen to which they are attached, form anoptionally substituted heterocycloalkyl group; and

where each optionally substituted group is unsubstituted orindependently substituted with one or more, such as one, two, or three,substituents independently selected from C₁-C₄ alkyl, aryl, heteroaryl,aryl-C₁-C₄ alkyl-, heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl, —OC₁-C₄alkyl, —OC₁-C₄ alkylphenyl, —C₁-C₄ alkyl-OH, —OC₁-C₄ haloalkyl, halo,—OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄ alkylphenyl),cyano, nitro, oxo (as a substitutent for cycloalkyl, heterocycloalkyl,or heteroaryl), —CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄ alkyl)(C₁-C₄alkyl), —CONH(C₁-C₄ alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl),—NHC(O)(phenyl), —N(C₁-C₄ alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl, —C(O)C₁-C₄ alkylphenyl, —C(O)C₁-C₄haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄ alkyl), —SO₂(phenyl),—SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄ alkyl), —SO₂NH(phenyl),—NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and —NHSO₂(C₁-C₄ haloalkyl).

In some embodiments, a substituted alkoxy group is “polyalkoxy” or —O—(optionally substituted alkylene)-(optionally substituted alkoxy), andincludes groups such as —OCH₂CH₂OCH₃, and residues of glycol ethers suchas polyethyleneglycol, and —O(CH₂CH₂O)_(x)CH₃, where x is an integer of2-20, such as 2-10, and for example, 2-5. Another substituted alkoxygroup is hydroxyalkoxy or —OCH₂(CH₂)_(y)OH, where y is an integer of1-10, such as 1-4.

The term “alkoxycarbonyl” refers to a group of the formula(alkoxy)(C═O)— attached through the carbonyl carbon wherein the alkoxygroup has the indicated number of carbon atoms. Thus a C₁-C₆alkoxycarbonyl group is an alkoxy group having from 1 to 6 carbon atomsattached through its oxygen to a carbonyl linker. “Lower alkoxycarbonyl”refers to an alkoxycarbonyl group wherein the alkoxy group is a loweralkoxy group.

The term “substituted alkoxycarbonyl” refers to the group (substitutedalkyl)-O—C(O)— wherein the group is attached to the parent structurethrough the carbonyl functionality and wherein substituted refers toalkyl wherein one or more (such as up to 5, for example, up to 3)hydrogen atoms are replaced by a substituent independently chosen from:

—R^(a), —OR^(b), optionally substituted amino (including —NR^(c)COR^(b),—NR^(c)CO₂R^(a), —NR^(c)CONR^(b)R^(c), —NR^(b)C(NR^(c))NR^(b)R^(c),—NR^(b)C(NCN)NR^(b)R^(c), and —NR^(c)SO₂R^(a)), halo, cyano, nitro, oxo(as a substitutent for cycloalkyl, heterocycloalkyl, and heteroaryl),optionally substituted acyl (such as —COR^(b)), optionally substitutedalkoxycarbonyl (such as —CO₂R^(b)), aminocarbonyl (such as—CON^(b)R^(c)), —OCOR^(b), —OCO₂R^(a), —OCOR^(b)R^(c), —OCONR^(b)R^(c),—OP(O)(OR^(b))OR^(c), sulfanyl (such as SR^(b)), sulfinyl (such as—SOR^(a)), and sulfonyl (such as —SO₂R^(a) and —SO₂NR^(b)R^(c)),

where R^(a) is chosen from optionally substituted C₁-C₆ alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, and optionally substituted heteroaryl;

R^(b) is chosen from H, optionally substituted C₁-C₆ alkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted aryl, and optionally substituted heteroaryl; and

R^(c) is independently chosen from hydrogen and optionally substitutedC₁-C₄ alkyl; or

R^(b) and R^(c), and the nitrogen to which they are attached, form anoptionally substituted heterocycloalkyl group; and

where each optionally substituted group is unsubstituted orindependently substituted with one or more, such as one, two, or three,substituents independently selected from C₁-C₄ alkyl, aryl, heteroaryl,aryl-C₁-C₄ alkyl-, heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl, —OC₁-C₄alkyl, —OC₁-C₄ alkylphenyl, —C₁-C₄ alkyl-OH, —OC₁-C₄ haloalkyl, halo,—OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄ alkylphenyl),cyano, nitro, oxo (as a substitutent for cycloalkyl, heterocycloalkyl,or heteroaryl), —CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄ alkyl)(C₁-C₄alkyl), —CONH(C₁-C₄ alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl),—NHC(O)(phenyl), —N(C₁-C₄ alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl, —C(O)C₁-C₄ alkylphenyl, —C(O)C₁-C₄haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄ alkyl), —SO₂(phenyl),—SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄ alkyl), —SO₂NH(phenyl),—NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and —NHSO₂(C₁-C₄ haloalkyl).

“Acyl” refers to the groups H—C(O)—, (alkyl)-C(O)—, (aryl)-C(O)—,(heteroaryl)-C(O)—, and (heterocycloalkyl)-C(O)—, wherein the group isattached to the parent structure through the carbonyl functionality, andwherein alkyl, aryl, heteroaryl, and heterocycloalkyl are optionallysubstituted as described herein. Examples include acetyl, benzoyl,propionyl, isobutyryl, t-butoxycarbonyl, benzyloxycarbonyl and the like.“Lower-acyl” refers to groups containing one to six carbons and“acyloxy” refers to the group O-acyl.

The term “amino” refers to the group —NH₂.

The term “substituted amino” refers to the group —NHR^(d) or—NR^(d)R^(e) wherein

R^(d) is chosen from hydroxy, optionally substituted alkoxy, optionallysubstituted alkyl, optionally substituted cycloalkyl, optionallysubstituted acyl, optionally substituted carbamimidoyl, aminocarbonyl,optionally substituted aryl, optionally substituted heteroaryl,optionally substituted heterocycloalkyl, optionally substitutedalkoxycarbonyl, sulfinyl and sulfonyl, and

R^(e) is chosen from optionally substituted alkyl, optionallysubstituted cycloalkyl, optionally substituted aryl, optionallysubstituted heteroaryl, and optionally substituted heterocycloalkyl, andwherein

substituted alkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroarylrefer respectively to alkyl, cycloalkyl, aryl, heterocycloalkyl, andheteroaryl wherein one or more (such as up to 5, for example, up to 3)hydrogen atoms are replaced by a substituent independently chosen from:

—R^(a), —OR^(b), optionally substituted amino (including —NR^(c)COR^(b),—NR^(c)CO₂R^(a), —NR^(c)CONR^(b)R^(c), —NR^(b)C(NR^(c))NR^(b)R^(c),—NR^(b)C(NCN)NR^(b)R^(c), and —NR^(c)SO₂R^(a)), halo, cyano, nitro, oxo(as a substitutent for cycloalkyl, heterocycloalkyl, and heteroaryl),optionally substituted acyl (such as —COR^(b)), optionally substitutedalkoxycarbonyl (such as —CO₂R^(b)), aminocarbonyl (such as—CONR^(b)R^(c)), —OCOR^(b), —OCO₂R^(a), —OCONR^(b)R^(c),—OCONR^(b)R^(c), —OP(O)(OR^(b))OR^(c), sulfanyl (such as SR^(b)),sulfinyl (such as —SOR^(a)), and sulfonyl (such as —SO₂R^(a) and—SO₂NR^(b)R^(c)), where

R^(a) is chosen from optionally substituted C₁-C₆ alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted aryl, and optionally substituted heteroaryl;

R^(b) is chosen from H, optionally substituted C₁-C₆ alkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted aryl, and optionally substituted heteroaryl; and

R^(c) is independently chosen from hydrogen and optionally substitutedC₁-C₄ alkyl; or R^(b) and R^(c), and the nitrogen to which they areattached, form an optionally substituted heterocycloalkyl group; andwhere each optionally substituted group is unsubstituted orindependently substituted with one or more, such as one, two, or three,substituents independently selected from C₁-C₄ alkyl, aryl, heteroaryl,aryl-C₁-C₄ alkyl-, heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl, —OC₁-C₄alkyl, —OC₁-C₄ alkylphenyl, —C₁-C₄ alkyl-OH, —OC₁-C₄ haloalkyl, halo,—OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄ alkylphenyl),cyano, nitro, oxo (as a substitutent for cycloalkyl, heterocycloalkyl,or heteroaryl), —CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄ alkyl)(C₁-C₄alkyl), —CONH(C₁-C₄ alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl),—NHC(O)(phenyl), —N(C₁-C₄ alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl, —C(O)C₁-C₄ alkylphenyl, —C(O)C₁-C₄haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄ alkyl), —SO₂(phenyl),—SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄ alkyl), —SO₂NH(phenyl),—NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and —NHSO₂(C₁-C₄ haloalkyl); andwherein

optionally substituted acyl, optionally substituted alkoxycarbonyl,sulfinyl and sulfonyl are as defined herein.

The term “substituted amino” also refers to N-oxides of the groups—NHR^(d), and NR^(d)R^(d) each as described above. N-oxides can beprepared by treatment of the corresponding amino group with, forexample, hydrogen peroxide or m-chloroperoxybenzoic acid. The personskilled in the art is familiar with reaction conditions for carrying outthe N-oxidation.

The term “aminocarbonyl” refers to the group —CONR^(b)R^(c), where R^(b)is chosen from H, optionally substituted C₁-C₆ alkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted aryl, and optionally substituted heteroaryl; and

R^(c) is independently chosen from hydrogen and optionally substitutedC₁-C₄ alkyl; or

R^(b) and R^(c) taken together with the nitrogen to which they arebound, form an optionally substituted 5- to 7-memberednitrogen-containing heterocycloalkyl which optionally includes 1 or 2additional heteroatoms selected from O, N, and S in the heterocycloalkylring;

where each substituted group is independently substituted with one ormore substituents independently selected from C₁-C₄ alkyl, aryl,heteroaryl, aryl-C₁-C₄ alkyl-, heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl,—OC₁-C₄ alkyl, —OC₁-C₄ alkylphenyl, —C₁-C₄ alkyl-OH, —OC₁-C₄ haloalkyl,halo, —OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄ alkyl),—NH(C₁-C₄ alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄alkylphenyl), cyano, nitro, oxo (as a substitutent for cycloalkyl,heterocycloalkyl, or heteroaryl), —CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄alkyl)(C₁-C₄ alkyl), —CONH(C₁-C₄ alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl),—NHC(O)(phenyl), —N(C₁-C₄ alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl, —C(O)C₁-C₄ alkylphenyl, —C(O)C₁-C₄haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄ alkyl), —SO₂(phenyl),—SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄ alkyl), —SO₂NH(phenyl),—NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and —NHSO₂(C₁-C₄ haloalkyl).

“Aryl” encompasses:

6-membered carbocyclic aromatic rings, for example, benzene;

bicyclic ring systems wherein at least one ring is carbocyclic andaromatic, for example, naphthalene, indane, and tetralin; and

tricyclic ring systems wherein at least one ring is carbocyclic andaromatic, for example, fluorene.

For example, aryl includes 6-membered carbocyclic aromatic rings fusedto a 5- to 7-membered heterocycloalkyl ring containing 1 or moreheteroatoms chosen from N, O, and S. For such fused, bicyclic ringsystems wherein only one of the rings is a carbocyclic aromatic ring,the point of attachment may be at the carbocyclic aromatic ring or theheterocycloalkyl ring. Bivalent radicals formed from substituted benzenederivatives and having the free valences at ring atoms are named assubstituted phenylene radicals. Bivalent radicals derived from univalentpolycyclic hydrocarbon radicals whose names end in “-yl” by removal ofone hydrogen atom from the carbon atom with the free valence are namedby adding “-idene” to the name of the corresponding univalent radical,e.g., a naphthyl group with two points of attachment is termednaphthylidene. Aryl, however, does not encompass or overlap in any waywith heteroaryl, separately defined below. Hence, if one or morecarbocyclic aromatic rings is fused with a heterocycloalkyl aromaticring, the resulting ring system is heteroaryl, not aryl, as definedherein.

“Aralkoxy” refers to the group —O-aralkyl. Similarly, “heteroaralkoxy”refers to the group —O-heteroaralkyl; “aryloxy” refers to —O-aryl; and“heteroaryloxy” refers to the group —O-heteroaryl.

“Aralkyl” refers to a residue in which an aryl moiety is attached to theparent structure via an alkyl residue. Examples include benzyl,phenethyl, phenylvinyl, phenylallyl and the like. “Heteroaralkyl” refersto a residue in which a heteroaryl moiety is attached to the parentstructure via an alkyl residue. Examples include furanylmethyl,pyridinylmethyl, pyrimidinylmethyl and the like.

“Halogen” or “halo” refers to fluorine, chlorine, bromine or iodine.Dihaloaryl, dihaloalkyl, trihaloaryl etc. refer to aryl and alkylsubstituted with a plurality of halogens, but not necessarily aplurality of the same halogen; thus 4-chloro-3-fluorophenyl is withinthe scope of dihaloaryl.

“Heteroaryl” encompasses:

5- to 7-membered aromatic, monocyclic rings containing one or more, forexample, from 1 to 4, or in certain embodiments, from 1 to 3,heteroatoms chosen from N, O, and S, with the remaining ring atoms beingcarbon;

bicyclic heterocycloalkyl rings containing one or more, for example,from 1 to 4, or in certain embodiments, from 1 to 3, heteroatoms chosenfrom N, O, and S, with the remaining ring atoms being carbon and whereinat least one heteroatom is present in an aromatic ring; and

tricyclic heterocycloalkyl rings containing one or more, for example,from 1 to 5, or in certain embodiments, from 1 to 4, heteroatoms chosenfrom N, O, and S, with the remaining ring atoms being carbon and whereinat least one heteroatom is present in an aromatic ring.

For example, heteroaryl includes a 5- to 7-membered heterocycloalkyl,aromatic ring fused to a 5- to 7-membered cycloalkyl or heterocycloalkylring. For such fused, bicyclic heteroaryl ring systems wherein only oneof the rings contains one or more heteroatoms, the point of attachmentmay be at either ring. When the total number of S and O atoms in theheteroaryl group exceeds 1, those heteroatoms are not adjacent to oneanother. In certain embodiments, the total number of S and O atoms inthe heteroaryl group is not more than 2. In certain embodiments, thetotal number of S and O atoms in the aromatic heterocycle is not morethan 1. Examples of heteroaryl groups include, but are not limited to,(as numbered from the linkage position assigned priority 1), 2-pyridyl,3-pyridyl, 4-pyridyl, 2,3-pyrazinyl, 3,4-pyrazinyl, 2,4-pyrimidinyl,3,5-pyrimidinyl, 2,3-pyrazolinyl, 2,4-imidazolinyl, isoxazolinyl,oxazolinyl, thiazolinyl, thiadiazolinyl, tetrazolyl, thienyl,benzothiophenyl, furanyl, benzofuranyl, benzoimidazolinyl, indolinyl,pyridazinyl, triazolyl, quinolinyl, pyrazolyl, and5,6,7,8-tetrahydroisoquinolinyl. Bivalent radicals derived fromunivalent heteroaryl radicals whose names end in “-yl” by removal of onehydrogen atom from the atom with the free valence are named by adding“-idene” to the name of the corresponding univalent radical, e.g., apyridyl group with two points of attachment is a pyridylidene.Heteroaryl does not encompass or overlap with aryl, cycloalkyl, orheterocycloalkyl, as defined herein

Substituted heteroaryl also includes ring systems substituted with oneor more oxide (—O⁻) substituents, such as pyridinyl N-oxides.

By “heterocycloalkyl” is meant a single, non-aromatic ring, usually with3 to 7 ring atoms, containing at least 2 carbon atoms in addition to 1-3heteroatoms independently selected from oxygen, sulfur, and nitrogen, aswell as combinations comprising at least one of the foregoingheteroatoms. The ring may be saturated or have one or more carbon-carbondouble bonds. Suitable heterocycloalkyl groups include, for example (asnumbered from the linkage position assigned priority 1), 2-pyrrolidinyl,2,4-imidazolidinyl, 2,3-pyrazolidinyl, 2-piperidyl, 3-piperidyl,4-piperidyl, and 2,5-piperizinyl. Morpholinyl groups are alsocontemplated, including 2-morpholinyl and 3-morpholinyl (numberedwherein the oxygen is assigned priority 1). Substituted heterocycloalkylalso includes ring systems substituted with one or more oxo (═O) oroxide (—O⁻) substituents, such as piperidinyl N-oxide,morpholinyl-N-oxide, 1-oxo-1-thiomorpholinyl and1,1-dioxo-1-thiomorpholinyl.

“Heterocycloalkyl” also includes bicyclic ring systems wherein onenon-aromatic ring, usually with 3 to 7 ring atoms, contains at least 2carbon atoms in addition to 1-3 heteroatoms independently selected fromoxygen, sulfur, and nitrogen, as well as combinations comprising atleast one of the foregoing heteroatoms; and the other ring, usually with3 to 7 ring atoms, optionally contains 1-3 heteroatom independentlyselected from oxygen, sulfur, and nitrogen and is not aromatic.

“Isomers” are different compounds that have the same molecular formula.“Stereoisomers” are isomers that differ only in the way the atoms arearranged in space. “Enantiomers” are a pair of stereoisomers that arenon-superimposable mirror images of each other. A 1:1 mixture of a pairof enantiomers is a “racemic” mixture. The term “(.±.)” is used todesignate a racemic mixture where appropriate. “Diastereoisomers” arestereoisomers that have at least two asymmetric atoms, but which are notmirror-images of each other. The absolute stereochemistry is specifiedaccording to the Cahn-Ingold-Prelog R-S system. When a compound is apure enantiomer the stereochemistry at each chiral carbon can bespecified by either R or S. Resolved compounds whose absoluteconfiguration is unknown can be designated (+) or (−) depending on thedirection (dextro- or levorotatory) which they rotate plane polarizedlight at the wavelength of the sodium D line. Certain of the compoundsdescribed herein contain one or more asymmetric centers and can thusgive rise to enantiomers, diastereomers, and other stereoisomeric formsthat can be defined, in terms of absolute stereochemistry, as (R)- or(S)-. The present invention is meant to include all such possibleisomers, including racemic mixtures, optically pure forms andintermediate mixtures. Optically active (R)- and (S)-isomers can beprepared using chiral synthons or chiral reagents, or resolved usingconventional techniques. When the compounds described herein containolefinic double bonds or other centers of geometric asymmetry, andunless specified otherwise, it is intended that the compounds includeboth E and Z geometric isomers.

“Tautomers” are structurally distinct isomers that interconvert bytautomerization. “Tautomerization” is a form of isomerization andincludes prototropic or proton-shift tautomerization, which isconsidered a subset of acid-base chemistry. “Prototropictautomerization” or “proton-shift tautomerization” involves themigration of a proton accompanied by changes in bond order, often theinterchange of a single bond with an adjacent double bond. Wheretautomerization is possible (e.g. in solution), a chemical equilibriumof tautomers can be reached. An example of tautomerization is keto-enoltautomerization. A specific example of keto-enol tautomerization is theinterconverision of pentane-2,4-dione and 4-hydroxypent-3-en-2-onetautomers. Another example of tautomerization is phenol-ketotautomerization. A specific example of phenol-keto tautomerization isthe interconversion of pyridin-4-ol and pyridin-4(1H)-one tautomers.Compounds of Formula I are tautomeric.

A leaving group or atom is any group or atom that will, under thereaction conditions, cleave from the starting material, thus promotingreaction at a specified site. Suitable examples of such groups unlessotherwise specified are halogen atoms, mesyloxy,p-nitrobenzensulphonyloxy and tosyloxy groups.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” includes any and all solvents, dispersion media,coatings, antibacterial and antifungal agents, isotonic and absorptiondelaying agents and the like. The use of such media and agents forpharmaceutically active substances is well known in the art. Exceptinsofar as any conventional media or agent is incompatible with theactive ingredient, its use in the therapeutic compositions iscontemplated. Supplementary active ingredients can also be incorporatedinto the compositions.

Protecting group has the meaning conventionally associated with it inorganic synthesis, i.e. a group that selectively blocks one or morereactive sites in a multifunctional compound such that a chemicalreaction can be carried out selectively on another unprotected reactivesite and such that the group can readily be removed after the selectivereaction is complete. A variety of protecting groups are disclosed, forexample, in T. H. Greene and P. G. M. Wuts, Protective Groups in OrganicSynthesis, Third Edition, John Wiley & Sons, New York (1999). Forexample, a hydroxy protected form is where at least one of the hydroxygroups present in a compound is protected with a hydroxy protectinggroup. Likewise, amines and other reactive groups may similarly beprotected.

The term “pharmaceutically acceptable salt” refers to salts that retainthe biological effectiveness and properties of the compounds describedherein and, which are not biologically or otherwise undesirable. In manycases, the compounds described herein are capable of forming acid and/orbase salts by virtue of the presence of amino and/or carboxyl groups orgroups similar thereto. Pharmaceutically acceptable acid addition saltscan be formed with inorganic acids and organic acids. Inorganic acidsfrom which salts can be derived include, for example, hydrochloric acid,hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and thelike. Organic acids from which salts can be derived include, forexample, acetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Pharmaceutically acceptable base additionsalts can be formed with inorganic and organic bases. Inorganic basesfrom which salts can be derived include, for example, sodium, potassium,lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese,aluminum, and the like. Organic bases from which salts can be derivedinclude, for example, primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines, basic ion exchange resins, and the like, specificallysuch as isopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, and ethanolamine. In some embodiments, thepharmaceutically acceptable base addition salt is chosen from ammonium,potassium, sodium, calcium, and magnesium salts.

The term “solvate” refers to a compound (e.g., a compound selected fromFormula I, or a pharmaceutically acceptable salt thereof) in physicalassociation with one or more molecules of a pharmaceutically acceptablesolvent. It will be understood that “a compound of Formula I”encompasses the compound of Formula I, and solvates of those compounds,as well as mixtures thereof.

The terms “substituted” alkyl, cycloalkyl, aryl, heterocycloalkyl, andheteroaryl, unless otherwise expressly defined, refer respectively toalkyl, cycloalkyl, aryl, heterocycloalkyl, and heteroaryl wherein one ormore (such as up to 5, for example, up to 3) hydrogen atoms are replacedby a substituent independently chosen from:

—R^(a), —OR^(b), optionally substituted amino (including —NR^(c)COR^(b),—NR^(c)CO₂R^(a), —NR^(c)CONR^(b)R^(c), —NR^(b)C(NR^(c))NR^(b)R^(c),—NR^(b)C(NCN)NR^(b)R^(c), and —NR^(c)SO₂R^(a)), halo, cyano, nitro, oxo(as a substitutent for cycloalkyl, heterocycloalkyl, and heteroaryl),optionally substituted acyl (such as —COR^(b)), optionally substitutedalkoxycarbonyl (such as —CO₂R^(b)), aminocarbonyl (such as—CONR^(b)R^(c)), —OCOR^(b), —OCO₂R^(a), —OCONR^(b)R^(c),—OCONR^(b)R^(c), —OP(O)(OR^(b))OR^(c), sulfanyl (such as SR^(b)),sulfinyl (such as —SOR^(a)), and sulfonyl (such as —SO₂R^(a) and—SO₂NR^(b)R^(c)), where

R^(a) is chosen from optionally substituted C₁-C₆ alkyl, optionallysubstituted cycloalkyl, optionally substituted heterocycloalkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted aryl, and optionally substituted heteroaryl;

R^(b) is chosen from hydrogen, optionally substituted C₁-C₆ alkyl,optionally substituted cycloalkyl, optionally substitutedheterocycloalkyl, optionally substituted aryl, and optionallysubstituted heteroaryl; and

R^(c) is independently chosen from hydrogen and optionally substitutedC₁-C₄ alkyl; or

R^(b) and R^(c), and the nitrogen to which they are attached, form anoptionally substituted heterocycloalkyl group; and

where each optionally substituted group is unsubstituted orindependently substituted with one or more, such as one, two, or three,substituents independently selected from C₁-C₄ alkyl, aryl, heteroaryl,aryl-C₁-C₄ alkyl-, heteroaryl-C₁-C₄ alkyl-, C₁-C₄ haloalkyl, —OC₁-C₄alkyl, —OC₁-C₄ alkylphenyl, —C₁-C₄ alkyl-OH, —OC₁-C₄ haloalkyl, halo,—OH, —NH₂, —C₁-C₄ alkyl-NH₂, —N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —NH(C₁-C₄alkyl), —N(C₁-C₄ alkyl)(C₁-C₄ alkylphenyl), —NH(C₁-C₄ alkylphenyl),cyano, nitro, oxo (as a substitutent for cycloalkyl orheterocycloalkyl), —CO₂H, —C(O)OC₁-C₄ alkyl, —CON(C₁-C₄ alkyl)(C₁-C₄alkyl), —CONH(C₁-C₄ alkyl), —CONH₂, —NHC(O)(C₁-C₄ alkyl),—NHC(O)(phenyl), —N(C₁-C₄ alkyl)C(O)(C₁-C₄ alkyl), —N(C₁-C₄alkyl)C(O)(phenyl), —C(O)C₁-C₄ alkyl, —C(O)C₁-C₄ alkylphenyl, —C(O)C₁-C₄haloalkyl, —OC(O)C₁-C₄ alkyl, —SO₂(C₁-C₄ alkyl), —SO₂(phenyl),—SO₂(C₁-C₄ haloalkyl), —SO₂NH₂, —SO₂NH(C₁-C₄ alkyl), —SO₂NH(phenyl),—NHSO₂(C₁-C₄ alkyl), —NHSO₂(phenyl), and —NHSO₂(C₁-C₄ haloalkyl).

The term “sulfanyl” refers to the groups: —S-(optionally substitutedalkyl), —S-(optionally substituted cycloalkyl), —S-(optionallysubstituted aryl), —S-(optionally substituted heteroaryl), and—S-(optionally substituted heterocycloalkyl).

The term “sulfinyl” refers to the groups: —S(O)—H, —S(O)-(optionallysubstituted alkyl), —S(O)-(optionally substituted cycloalkyl),—S(O)-(optionally substituted amino), —S(O)-(optionally substitutedaryl), —S(O)-(optionally substituted heteroaryl), and —S(O)-(optionallysubstituted heterocycloalkyl).

The term “sulfonyl” refers to the groups: —S(O₂)—H, —S(O₂)-(optionallysubstituted alkyl), —S(O₂)-(optionally substituted cycloalkyl),—S(O₂)-(optionally substituted amino), —S(O₂)-(optionally substitutedaryl), —S(O₂)-(optionally substituted heteroaryl), and—S(O₂)-(optionally substituted heterocycloalkyl).

The term “therapeutically effective amount” or “effective amount” refersto that amount of a compound selected from Formula I that is sufficientto effect treatment, as defined below, when administered to a mammal inneed of such treatment. The therapeutically effective amount will varydepending upon the subject and disease condition being treated, theweight and age of the subject, the severity of the disease condition,the particular compound selected from Formula I, the dosing regimen tobe followed, timing of administration, the manner of administration andthe like, all of which can readily be determined by one of ordinaryskill in the art.

“ATPase” refers to an enzyme that hydrolyzes ATP. ATPases includeproteins comprising molecular motors such as the myosins.

As used herein, “frailty” is a syndrome characterized by meeting threeof the of the following five attributes: unintentional weight loss,muscle weakness, slow walking speed, exhaustion, and low physicalactivity.

As used herein, “cachexia” means a metabolic defect often associatedwith cancer that is characterized by progressive weight loss due to thedeletion of adipose tissue and skeletal muscle.

As used herein, “muscle spasm” means an involuntary contraction of amuscle. Muscle spasms may lead to cramps.

As used herein, “post-surgical muscle weakness” refers to a reduction inthe strength of one or more muscles following surgical procedure.Weakness may be generalized (i.e. total body weakness) or localized to aspecific area, side of the body, limb, or muscle.

As used herein, “post-traumatic muscle weakness” refers to a reductionin the strength of one or more muscles following a traumatic episode(e.g. bodily injury). Weakness may be generalized (i.e. total bodyweakness) or localized to a specific area, side of the body, limb, ormuscle.

As used herein, “neuromuscular disease” means any disease that affectsany part of the nerve and muscle. Neuromuscular disease encompassescritical illness polyneuropathy, prolonged neuromuscular blockade, acutemyopathy as well as acute inflammatory demyelinatingpolyradiculoneuropathy, amyotrophic lateral sclerosis (ALS), autonomicneuropathy, Charcot-Marie-Tooth disease and other hereditary motor andsensory neuropathies, chronic inflammatory demyelinatingpolyradiculoneuropathy, dermatomyositis/polymyositis, diabeticneuropathy, dystrophinopathies, endocrine myopathies, focal muscularatrophies, hemifacial spasm, hereditary neuropathies of theCharcot-Marie-Tooth disease type, inclusion body myositis, Kennedydisease, Lambert-Eaton myasthenic syndrome, muscular dystrophy (e.g.,limb-girdle, Duchenne, Becker, myotonic, facioscapulohumeral, etc.),metabolic myopathies, metabolic neuropathy, multifocal motor neuropathywith conduction blocks, myasthenia gravis, neuropathy of FriedreichAtaxia, neuropathy of leprosy, nutritional neuropathy, periodicparalyses, primary lateral sclerosis, restrictive lung disease,sarcoidosis and neuropathy, Schwartz-Jampel Syndrome, spinal muscleatrophy, stiff person syndrome, thyroid disease, traumatic peripheralnerve lesions, vasculitic neuropathy, among others.

As used herein “obesity” means having a body mass index (BMI) greaterthan or equal to 30 kg/m². BMI is defined as weight (kg) divided byheight (m²). Obesity encompasses hyperplastic obesity, an increase inthe number of fat cells, and hypertrophic obesity, an increase in thesize of the fat cells. Overweight is defined as having a BMI from 25 upto 30 kg/m²; obesity as a BMI greater than or equal to 30 kg/m², asstated above, and severe (or morbid) obesity is defined as a BMI greaterthan or quality to 40 kg/m².

As used herein, “sarcopenia” means a loss of skeletal muscle mass,quality, and strength. Often sarcopenia is attributed to ageing, but isalso associated with HIV infection. Sarcopenia may lead to frailty, forexample, in the elderly.

As used herein, “wasting syndrome” means a condition characterized byinvoluntary weight loss associated with chronic fever and diarrhea. Insome instances, patients with wasting syndrome lose 10% of baseline bodyweight within one month.

Compounds of Formula I also include crystalline and amorphous forms ofthose compounds, including, for example, polymorphs, pseudopolymorphs,solvates, hydrates, unsolvated polymorphs (including anhydrates),conformational polymorphs, and amorphous forms of the compounds, as wellas mixtures thereof. “Crystalline form,” “polymorph,” and “novel form”may be used interchangeably herein, and are meant to include allcrystalline and amorphous forms of the compound, including, for example,polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs(including anhydrates), conformational polymorphs, and amorphous forms,as well as mixtures thereof, unless a particular crystalline oramorphous form is referred to.

Chemical entities include, but are not limited to, compounds of FormulaI and all pharmaceutically acceptable forms thereof. Pharmaceuticallyacceptable forms of the compounds recited herein includepharmaceutically acceptable salts, chelates, non-covalent complexes,prodrugs, and mixtures thereof. In certain embodiments, the compoundsdescribed herein are in the form of pharmaceutically acceptable salts.Hence, the terms “chemical entity” and “chemical entities” alsoencompass pharmaceutically acceptable salts, chelates, non-covalentcomplexes, prodrugs, and mixtures.

“Pharmaceutically acceptable salts” include, but are not limited tosalts with inorganic acids, such as hydrochlorate, phosphate,diphosphate, hydrobromate, sulfate, sulfinate, nitrate, and like salts;as well as salts with an organic acid, such as malate, maleate,fumarate, tartrate, succinate, citrate, acetate, lactate,methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate,salicylate, stearate, and alkanoate such as acetate, HOOC—(CH₂)_(n)—COOHwhere n ranges from 0 to 4, and like salts. Similarly, pharmaceuticallyacceptable cations include, but are not limited to sodium, potassium,calcium, aluminum, lithium, and ammonium.

In addition, if the compound of Formula I is obtained as an acidaddition salt, the free base can be obtained by basifying a solution ofthe acid salt. Conversely, if the product is a free base, an additionsalt, particularly a pharmaceutically acceptable addition salt, may beproduced by dissolving the free base in a suitable organic solvent andtreating the solution with an acid, in accordance with conventionalprocedures for preparing acid addition salts from base compounds. Thoseskilled in the art will recognize various synthetic methodologies thatmay be used to prepare non-toxic pharmaceutically acceptable additionsalts.

As noted above, prodrugs also fall within the scope of chemicalentities, for example, ester or amide derivatives of the compoundsselected from Formula I. The term “prodrug” includes any compound thatbecomes a compound of Formula I when administered to a patient, e.g.,upon metabolic processing of the prodrug. Examples of prodrugs include,but are not limited to, acetate, formate, benzoate, and like derivativesof functional groups (such as alcohol or amine groups) in the compoundsselected from Formula I.

The term “chelate” refers to the chemical entity formed by thecoordination of a compound to a metal ion at two (or more) points.

The term “non-covalent complex” refers to the chemical entity formed bythe interaction of a compound and another molecule wherein a covalentbond is not formed between the compound and the molecule. For example,complexation can occur through van der Waals interactions, hydrogenbonding, and electrostatic interactions (also called ionic bonding).

The term “active agent” is used to indicate a chemical entity which hasbiological activity. In certain embodiments, an “active agent” is acompound having pharmaceutical utility.

The term “therapeutically effective amount” of a chemical entity meansan amount effective, when administered to a human or non-human patient,to treat a disease, e.g., a therapeutically effective amount may be anamount sufficient to treat a disease or disorder responsive to myosinactivation. The therapeutically effective amount may be ascertainedexperimentally, for example by assaying blood concentration of thechemical entity, or theoretically, by calculating bioavailability.

By “significant” is meant any detectable change that is statisticallysignificant in a standard parametric test of statistical significancesuch as Student's T-test, where p<0.05.

“Patient” refers to an animal, such as a mammal, for example a human,that has been or will be the object of treatment, observation orexperiment. The methods described herein can be useful in both humantherapy and veterinary applications. In some embodiments, the patient isa mammal, and in some embodiments, the patient is human.

“Treatment” or “treating” means any treatment of a disease in a patient,including:

-   -   (a) preventing the disease, that is, causing the clinical        symptoms of the disease not to develop;    -   (b) inhibiting the disease;    -   (c) slowing or arresting the development of clinical symptoms;        and/or    -   (d) relieving the disease, that is, causing the regression of        clinical symptoms.

As used herein, “modulation” refers to a change in one or more ofskeletal myosin, skeletal actin, skeletal tropomyosin, skeletal troponinC, skeletal troponin I, skeletal troponin T, and skeletal muscle,including fragments and isoforms thereof, as well as the skeletalsarcomere as a direct or indirect response to the presence of at leastone chemical entity described herein, relative to the activity of themyosin or sarcomere in the absence of the compound. The change may be anincrease in activity (potentiation) or a decrease in activity(inhibition), and may be due to the direct interaction of the compoundwith myosin or the sarcomere, or due to the interaction of the compoundwith one or more other factors that in turn effect one or more ofskeletal myosin, skeletal actin, skeletal tropomyosin, skeletal troponinC, skeletal troponin I, skeletal troponin T, and skeletal muscle,including fragments and isoforms thereof, as well as the skeletalsarcomere, for example, through sensitization of the skeletal myosin orthe sarcomere to contraction at lower Ca²⁺ concentrations.

Provided is at least one chemical entity chosen from compounds ofFormula I:

and pharmaceutically acceptable salts and tautomers thereof, wherein

R³ is selected from optionally substituted alkyl, optionally substitutedcycloalkyl, optionally substituted aryl, optionally substitutedheteroaryl, and optionally substituted heterocycloalkyl;

R⁵ is selected from halo, hydroxy, optionally substituted alkyl,optionally substituted amino, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted aryl, optionally substitutedcycloalkyl, optionally substituted heteroaryl, optionally substitutedheterocycloalkyl, optionally substituted alkoxy, sulfonyl, sulfanyl, andsulfinyl;

R⁶ is selected from hydrogen, halo, hydroxy, lower alkyl, and lowerhaloalkyl; and

R⁷ is selected from hydrogen, optionally substituted acyl, optionallysubstituted alkyl, optionally substituted aryl, optionally substitutedcycloalkyl, optionally substituted heteroaryl, optionally substitutedaminocarbonyl, sulfonyl, sulfanyl, sulfinyl, carboxy, optionallysubstituted alkoxycarbonyl, hydroxy, and cyano.

In some embodiments, R³ is selected from optionally substitutedcycloalkyl and optionally substituted lower alkyl. In some embodiments,R³ is lower alkyl optionally substituted with one or more groupsselected from optionally substituted phenyl, hydroxy, optionallysubstituted alkoxy, optionally substituted amino and optionallysubstituted heterocycloalkyl. In some embodiments, R³ is lower alkyloptionally substituted with one or more groups selected from hydroxy,optionally substituted alkoxy, and optionally substituted amino. In someembodiments, R³ is selected from lower alkyl and lower alkyl substitutedwith hydroxyl. In some embodiments, R³ is lower alkyl substituted withhydroxyl. In some embodiments, R³ is lower alkyl. In some embodiments,R³ is pentyl. In some embodiments, R³ is

In some embodiments, R⁵ is selected from halo, optionally substitutedamino, optionally substituted lower alkyl, optionally substitutedalkenyl, and optionally substituted alkynyl. In some embodiments, R⁵ isselected from prop-1-enyl, ethynyl, halo, vinyl, 1-methylvinyl, amino,alkylamino, and (dialkyl)amino. In some embodiments, R⁵ is selected fromethynyl, prop-1-enyl and 1-methylvinyl.

In some embodiments, R⁶ is hydrogen.

In some embodiments, R⁷ is selected from hydrogen and lower alkyl. Insome embodiments, R⁷ is hydrogen.

In some embodiments, the compound of Formula I is chosen from

The chemical entities described herein can be synthesized utilizingtechniques well known in the art, e.g., as illustrated below withreference to the Reaction Schemes.

Unless specified to the contrary, the reactions described herein takeplace at atmospheric pressure, generally within a temperature range from−10° C. to 200° C. Further, except as employed in the Examples or asotherwise specified, reaction times and conditions are intended to beapproximate, e.g., taking place at about atmospheric pressure within atemperature range of about −10° C. to about 110° C. over a period ofabout 1 to about 24 hours; reactions left to run overnight average aperiod of about 16 hours.

The terms “solvent,” “organic solvent,” and “inert solvent” each mean asolvent inert under the conditions of the reaction being described inconjunction therewith [including, for example, benzene, toluene,acetonitrile, tetrahydrofuran (“THF”), dimethylformamide (“DMF”),chloroform, methylene chloride (or dichloromethane), diethyl ether,methanol, N-methylpyrrolidone (“NMP”), pyridine and the like]. Unlessspecified to the contrary, the solvents used in the reactions describedherein are inert organic solvents. Unless specified to the contrary, foreach gram of the limiting reagent, one cc (or mL) of solvent constitutesa volume equivalent

Isolation and purification of the chemical entities and intermediatesdescribed herein can be effected, if desired, by any suitable separationor purification procedure such as, for example, filtration, extraction,crystallization, column chromatography, thin-layer chromatography orthick-layer chromatography, or a combination of these procedures.Specific illustrations of suitable separation and isolation procedurescan be had by reference to the examples hereinbelow. However, otherequivalent separation or isolation procedures can also be used.

When desired, the (R)- and (S)-isomers may be resolved by methods knownto those skilled in the art, for example by formation ofdiastereoisomeric salts or complexes which may be separated, forexample, by crystallization; via formation of diastereoisomericderivatives which may be separated, for example, by crystallization,gas-liquid or liquid chromatography; selective reaction of oneenantiomer with an enantiomer-specific reagent, for example enzymaticoxidation or reduction, followed by separation of the modified andunmodified enantiomers; or gas-liquid or liquid chromatography in achiral environment, for example on a chiral support, such as silica witha bound chiral ligand or in the presence of a chiral solvent.Alternatively, a specific enantiomer may be synthesized by asymmetricsynthesis using optically active reagents, substrates, catalysts orsolvents, or by converting one enantiomer to the other by asymmetrictransformation.

Many of the optionally substituted starting compounds and otherreactants are commercially available, e.g., from Aldrich ChemicalCompany (Milwaukee, Wis.) or can be readily prepared by those skilled inthe art using commonly employed synthetic methodology.

Referring to Reaction Scheme 1, step 1, a compound of Formula 101 isstirred with an excess of base, such as about 2 equiv. of K₂CO₃, in asolvent, such as acetonitrile, and amine of formula NH₂R³. The product,a compound of Formula 102, is isolated and optionally purified.

Referring to Reaction Scheme 1, step 2, a compound of Formula 102 isdissolved in solvent, such as MeOH, and combined with a catalytic amountof Pd/C, such as about 0.05 equiv. The resulting mixture is hydrogenatedusing, for example, a pressure of about 50 psi. The intermediate amineis isolated and optionally purified. The amine is heated with an excess,such as about 3 equiv., of carbonyl diimidazole, in an inert solvent.The product, a compound of Formula 103, is isolated and optionallypurified.

A racemic mixture is optionally placed on a chromatography column andseparated into (R)- and (S)-enantiomers.

The compounds described herein are optionally contacted with apharmaceutically acceptable acid to form the corresponding acid additionsalts.

Pharmaceutically acceptable acid addition salts of compounds of FormulaI are optionally contacted with a base to form the corresponding freebase.

The chemical entities described herein modulate one or more of skeletalmyosin, skeletal actin, skeletal tropomyosin, skeletal troponin C,skeletal troponin I, skeletal troponin T, and skeletal muscle, includingfragments and isoforms thereof, as well as the skeletal sarcomere, andare useful to bind to, inhibit and/or potentiate the activity thereof.As used in this context, “modulate” means either increasing ordecreasing myosin activity, whereas “potentiate” means to increaseactivity and “inhibit” means to decrease activity.

The chemical entities, pharmaceutical compositions and methods describedherein are used to treat obesity, sarcopenia, wasting syndrome, frailty,cachexia, muscle spasm, post-surgical and post-traumatic muscleweakness, neuromuscular disease, and other indications in a mammal.

Methods to identify the chemical entities as binding to a protein or asa modulator of the binding characteristics or biological activity of aprotein are described in, for example, U.S. Pat. No. 6,410,254 and U.S.patent application Ser. No. 10/987,165.

For example, test compounds can be assayed in a highly parallel fashionby using multiwell plates by placing the compounds either individuallyin wells or testing them in mixtures. Assay components including thetarget protein complex, coupling enzymes and substrates, and ATP canthen be added to the wells and the absorbance or fluorescence of eachwell of the plate can be measured with a plate reader.

In some embodiments, the method uses a 384 well plate format and a 25 μLreaction volume. A pyruvate kinase/lactate dehydrogenase coupled enzymesystem (Huang T G and Hackney D D. (1994) J Biol Chem 269(23):16493-501)can be used to measure the rate of ATP hydrolysis in each Well. As willbe appreciated by those in the art, the assay components are added inbuffers and reagents. The incubation periods can be optimized to giveadequate detection signals over the background. The assay can be done inreal time giving the kinetics of ATP hydrolysis which increases thesignal to noise ratio of the assay.

The compounds can be further tested using skinned muscle fiberpreparations. Such assays are known in the art. See, e.g., Cheung et al.(2002) Nature Cell Biol. 4:83 and U.S. Patent Publication No.20020006962.

The chemical entities described herein are administered at atherapeutically effective dosage, e.g., a dosage sufficient to providetreatment for the disease states previously described. While humandosage levels have yet to be optimized for the chemical entitiesdescribed herein, generally, a daily dose ranges from about 0.05 to 100mg/kg of body weight; in certain embodiments, from about 0.10 to 10.0mg/kg of body weight, and in certain embodiments, from about 0.15 to 1.0mg/kg of body weight. Thus, for administration to a 70 kg person, incertain embodiments, the dosage range would be about from 3.5 to 7000 mgper day; in certain embodiments, about from 7.0 to 700.0 mg per day, andin certain embodiments, about from 10.0 to 100.0 mg per day. The amountof the chemical entity administered will, of course, be dependent on thesubject and disease state being treated, the severity of the affliction,the manner and schedule of administration and the judgment of theprescribing physician; for example, a likely dose range for oraladministration would be from about 70 to 700 mg per day, whereas forintravenous administration a likely dose range would be from about 70 to700 mg per day depending on compound pharmacokinetics.

Administration of the chemical entities described herein can be via anyof the accepted modes of administration for agents that serve similarutilities including, but not limited to, orally, sublingually,subcutaneously, intravenously, intranasally, topically, transdermally,intraperitoneally, intramuscularly, intrapulmonarilly, vaginally,rectally, or intraocularly. In some embodiments, oral or parenteraladministration is used.

Pharmaceutically acceptable compositions include solid, semi-solid,liquid and aerosol dosage forms, such as, e.g., tablets, capsules,powders, liquids, suspensions, suppositories, aerosols or the like. Thechemical entities can also be administered in sustained or controlledrelease dosage forms, including depot injections, osmotic pumps, pills,transdermal (including electrotransport) patches, and the like, forprolonged and/or timed, pulsed administration at a predetermined rate.In certain embodiments, the compositions are provided in unit dosageforms suitable for single administration of a precise dose.

The chemical entities described herein can be administered either aloneor more typically in combination with a conventional pharmaceuticalcarrier, excipient or the like (e.g., mannitol, lactose, starch,magnesium stearate, sodium saccharine, talcum, cellulose, sodiumcrosscarmellose, glucose, gelatin, sucrose, magnesium carbonate, and thelike). If desired, the pharmaceutical composition can also contain minoramounts of nontoxic auxiliary substances such as wetting agents,emulsifying agents, solubilizing agents, pH buffering agents and thelike (e.g., sodium acetate, sodium citrate, cyclodextrine derivatives,sorbitan monolaurate, triethanolamine acetate, triethanolamine oleate,and the like). Generally, depending on the intended mode ofadministration, the pharmaceutical composition will contain about 0.005%to 95%; in certain embodiments, about 0.5% to 50% by weight of achemical entity. Actual methods of preparing such dosage forms areknown, or will be apparent, to those skilled in this art; for example,see Remington's Pharmaceutical Sciences, Mack Publishing Company,Easton, Pa.

In addition, the chemical entities described herein can beco-administered with, and the pharmaceutical compositions can include,other medicinal agents, pharmaceutical agents, adjuvants, and the like.Suitable medicinal and pharmaceutical agents include modulators of oneor more of skeletal myosin, skeletal actin, skeletal tropomyosin,skeletal troponin C, skeletal troponin I, skeletal troponin T, andskeletal muscle, including fragments and isoforms thereof, and theskeletal sarcomere and other suitable therapeutic agents useful in thetreatment of the aforementioned disorders including: anti-obesityagents, anti-sarcopenia agents, anti-wasting syndrome agents,anti-frailty agents, anti-cachexia agents, anti-muscle spasm agents,agents against post-surgical and post-traumatic muscle weakness, andanti-neuromuscular disease agents, as well as the agents described inU.S. Patent Application No. 2005/0197367.

Suitable additional medicinal and pharmaceutical agents include, forexample: orlistat, sibramine, diethylpropion, phentermine,benzaphetamine, phendimetrazine, estrogen, estradiol, levonorgestrel,norethindrone acetate, estradiol valerate, ethinyl estradiol,norgestimate, conjugated estrogens, esterified estrogens,medroxyprogesterone acetate, testosterone, insulin-derived growthfactor, human growth hormone, riluzole, cannabidiol, prednisone,albuterol, non-steroidal anti-inflammatory drugs, and botulinum toxin.

Other suitable medicinal and pharmaceutical agents include TRH,diethylstilbestrol, theophylline, enkephalins, E series prostaglandins,compounds disclosed in U.S. Pat. No. 3,239,345 (e.g., zeranol),compounds disclosed in U.S. Pat. No. 4,036,979 (e.g., sulbenox),peptides disclosed in U.S. Pat. No. 4,411,890 growth hormonesecretagogues such as GHRP-6, GHRP-1 (disclosed in U.S. Pat. No.4,411,890 and publications WO 89/07110 and WO 89/07111), GHRP-2(disclosed in WO 93/04081), NN703 (Novo Nordisk), LY444711 (Lilly),MK-677 (Merck), CP424391 (Pfizer) and B-HT920, growth hormone releasingfactor and its analogs, growth hormone and its analogs and somatomedinsincluding IGF-1 and IGF-2, alpha-adrenergic agonists, such as clonidineor serotonin 5-HT_(D) agonists, such as sumatriptan, agents whichinhibit somatostatin or its release, such as physostigmine,pyridostigmine, parathyroid hormone, PTH(1-34), and bisphosphonates,such as MK-217 (alendronate).

Still other suitable medicinal and pharmaceutical agents includeestrogen, testosterone, selective estrogen receptor modulators, such astamoxifen or raloxifene, other androgen receptor modulators, such asthose disclosed in Edwards, J. P. et. al., Bio. Med. Chem. Let., 9,1003-1008 (1999) and Hamann, L. G. et. al., J. Med. Chem., 42, 210-212(1999), and progesterone receptor agonists (“PRA”), such aslevonorgestrel, medroxyprogesterone acetate (MPA).

Still other suitable medicinal and pharmaceutical agents include aP2inhibitors, such as those disclosed in U.S. Ser. No. 09/519,079 filedMar. 6, 2000, PPAR gamma antagonists, PPAR delta agonists, beta 3adrenergic agonists, such as AJ9677 (Takeda/Dainippon), L750355 (Merck),or CP331648 (Pfizer), other beta 3 agonists as disclosed in U.S. Pat.Nos. 5,541,204, 5,770,615, 5,491,134, 5,776,983 and 5,488,064, a lipaseinhibitor, such as orlistat or ATL-962 (Alizyme), a serotonin (anddopamine) reuptake inhibitor, such as sibutramine, topiramate (Johnson &Johnson) or axokine (Regeneron), a thyroid receptor beta drug, such as athyroid receptor ligand as disclosed in WO 97/21993, WO 99/00353, andGB98/284425, and anorectic agents, such as dexamphetamine, phentermine,phenylpropanolamine or mazindol.

Still other suitable medicinal and pharmaceutical agents include HIV andAIDS therapies, such as indinavir sulfate, saquinavir, saquinavirmesylate, ritonavir, lamivudine, zidovudine, lamivudine/zidovudinecombinations, zalcitabine, didanosine, stavudine, and megestrol acetate.

Still other suitable medicinal and pharmaceutical agents includeantiresorptive agents, hormone replacement therapies, vitamin Danalogues, elemental calcium and calcium supplements, cathepsin Kinhibitors, MMP inhibitors, vitronectin receptor antagonists, SrcSH.sub.2 antagonists, vacular—H⁺-ATPase inhibitors, ipriflavone,fluoride, Tibo lone, pro stanoids, 17-beta hydroxysteroid dehydrogenaseinhibitors and Src kinase inhibitors.

The above other therapeutic agents, when employed in combination withthe chemical entities described herein, may be used, for example, inthose amounts indicated in the Physicians' Desk Reference (PDR) or asotherwise determined by one of ordinary skill in the art.

In certain embodiments, the compositions will take the form of a pill ortablet and thus the composition will contain, along with the activeingredient, a diluent such as lactose, sucrose, dicalcium phosphate, orthe like; a lubricant such as magnesium stearate or the like; and abinder such as starch, gum acacia, polyvinylpyrrolidine, gelatin,cellulose, cellulose derivatives or the like. In another solid dosageform, a powder, marume, solution or suspension (e.g., in propylenecarbonate, vegetable oils or triglycerides) is encapsulated in a gelatincapsule.

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc. at least one chemical entityand optional pharmaceutical adjuvants in a carrier (e.g., water, saline,aqueous dextrose, glycerol, glycols, ethanol or the like) to form asolution or suspension. Injectables can be prepared in conventionalforms, either as liquid solutions or suspensions, as emulsions, or insolid forms suitable for dissolution or suspension in liquid prior toinjection. The percentage of chemical entities contained in suchparenteral compositions is highly dependent on the specific naturethereof, as well as the activity of the chemical entities and the needsof the subject. However, percentages of active ingredient of 0.01% to10% in solution are employable, and will be higher if the composition isa solid which will be subsequently diluted to the above percentages. Incertain embodiments, the composition will comprise from about 0.2 to 2%of the active agent in solution.

Pharmaceutical compositions of the chemical entities described hereinmay also be administered to the respiratory tract as an aerosol orsolution for a nebulizer, or as a microfine powder for insufflation,alone or in combination with an inert carrier such as lactose. In such acase, the particles of the pharmaceutical composition have diameters ofless than 50 microns, in certain embodiments, less than 10 microns.

EXAMPLES

The following examples serve to more fully describe the manner of usingthe above-described invention. It is understood that these examples inno way serve to limit the true scope of this invention, but rather arepresented for illustrative purposes.

Example I Preparation of5-(dimethylamino)-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol

5-(dimethylamino)-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol

6-Chloro-3-nitro-N-(pentan-3-yl)pyridin-2-amine. To a 0° C. mixture of2,6-dichloro-3-nitropyridine (12 g, 62 mmol) and potassium carbonate(17.1 g, 1.24 mmol) in acetonitrile (200 mL) was added 3-amino pentaneby syringe over 30 minutes. After stirring overnight at roomtemperature, the reaction was concentrated in vacuo. The residue wasdissolved in EtOAc and water and the organic layer was washed twice withwater and once with brine, dried over sodium sulfate, filtered andconcentrated in vacuo. Silica gel chromatography (5%-35% EtOAc/Hexanes)provided the title compound as a yellow solid (7.3 g, 48%). m/z=244.1(M+H)+

N⁶,N⁶-Dimethyl-3-nitro-N²-(pentan-3-yl)pyridine-2,6-diamine. To aroom-temperature mixture containing6-chloro-3-nitro-N-(pentan-3-yl)pyridin-2-amine (250 mg, 0.90 mmol) andpotassium carbonate (250 mg, 1.8 mmol) in acetonitrile (2 mL) was addeddimethylamine (1.35 mL, 1.0 M, 1.35 mmol) by syringe. After 3 h, thereaction mixture was concentrated in vacuo and then re-dissolved inEtOAc and water. The organic layer was washed twice with water and oncewith brine, dried over sodium sulfate, filtered and concentrated Invacuo. Silica gel chromatography (5%-20% EtOAc/Hexanes) provided thetitle compound as a yellow solid (119 mg, 52%). m/z=253.2 (M+H)+

5-(dimethylamino)-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol. To asolution of N⁶,N⁶-dimethyl-3-nitro-N²-(pentan-3-yl)pyridine-2,6-diamine(100 mg, 0.40 mmol) in MeOH (4 mL) was added a catalytic amount of Pd/C.The mixture was purged with hydrogen from a balloon fitted with a needlefor 1 h. The resulting mixture was filtered through a pad of celite andconcentrated in vacuo. The solid was then dissolved in THF (2 mL) andCDI (100 mg, 1.17 mmol) was added as a solid. After heating for 3 h, thereaction was cooled to room temperature. The excess CDI was quenched bythe careful addition of water. The mixture was diluted with EtOAc andwashed three times with water and once with brine. The solution was thendried over sodium sulfate, filtered and concentrated in vacuo. Silicagel chromatography (5%-25% EtOAc/hexanes) provided the title compound asa white solid (57 mg, 57%). m/z=249.2 (M+H)+

(CK-2018279) 3-(pentan-3-yl)-5-vinyl-3H-imidazo[4,5-b]pyridin-2-ol

6-Chloro-N²-(pentan-3-yl)pyridine-2,3-diamine. To a room-temperaturesolution containing 6-chloro-3-nitro-N-(pentan-3-yl)pyridin-2-amine (1.8g, 7.4 mmol) in MeOH (10 mL) and EtOAc (10 mL) was added SnCl₂ dihydrate(10.0 g, 44.3 mmol). After 3 h, the reaction mixture was concentrated invacuo and then re-dissolved in EtOAc and water. After stirringovernight, the layers were separated and the organic layer was driedover sodium sulfate, filtered, and concentrated in vacuo. Silica gelchromatography (25%-50% EtOAc/Hexanes) provided the title compound as awhite solid (940 mg, 57%). m/z=223.2 (M+H)+

5-Chloro-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol. To a solution of6-chloro-N²-(pentan-3-yl)pyridine-2,3-diamine (936 mg, 4.37 mmol) in THF(10 mL) and CDI (1.1 g, 12.9 mmol) was added as a solid. After heatingfor 3 h, the reaction was cooled to room temperature. The excess CDI wasquenched by the careful addition of water. The mixture was diluted withEtOAc and washed three times with water and once with brine. Thesolution was then dried over sodium sulfate, filtered and concentratedin vacuo. Silica gel chromatography (5%-40% EtOAc/hexanes) provided thetitle compound as a light purple solid (682 mg, 65%). m/z=240.0 (M+H)+

5-Vinyl-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol. To a microwavevial were added 5-chloro-3-(pentan-3-yl)-3H-imidazo[4,5-b]pyridin-2-ol(75 mg, 0.26 mmol), trivinylboroxine pyridine complex (30 mg, 0.12 mmol)and (DPPF)PdCl₂ (20 mg, 0.03 mmol). The vial was fitted with a septumand purged with nitrogen for 5 min. Dioxane (1.5 mL) and degassed 2 MK₂CO₃ (0.5 mL) were added by syringe, and the vial was capped and heatedin the microwave at 125° C. for 15 minutes. The mixture was diluted withEtOAc and washed three times with water and once with brine. The organiclayer was then dried over sodium sulfate, filtered and concentrated invacuo. Silica gel chromatography (25%-75% EtOAc/hexanes) provided thetitle compound as a yellow oil (26 mg, 43%). m/z=230.0 (M−H)−

Example 2 Preparation of Sarcomeric Proteins from Skeletal Muscle

Actin was purified by first preparing an ether powder of cardiac muscle(Zot H G and Potter J D. (1981) Preparative Biochemistry 11:381-395) asdescribed below. Subsequently, actin was cycled between the filamentousand soluble state through rounds of centrifugation and dialysis (SpudichJ A and Watt S. (1971) J. Biol. Chem. 246:4866-4871). It was stored inthe filamentous state at 4° C.

Tropomyosin was extracted from the ether powder and separated from theother proteins based on pH dependent precipitations followed bysuccessive ammonium sulfate cuts at 53% and 65% (Smillie L B. (1981)Methods Enzymol 85 Pt B:234-41). The troponins were isolated as anintact complex of TnC, TnT, and TnI. Ether powder is extracted in a highsalt buffer. Successive ammonium sulfate cuts of 30% and 45% were done;the precipitate was solubilized by dialysis into a low salt buffer andthen further purified on a DEAE Toyopearl column with a 25-350 mM KClgradient. There was no measurable ATPase in any of the components exceptfor myosin which naturally had a very low basal ATPase in the absence ofactin.

Just prior to screening, the actin, tropomyosin, and troponin complexare mixed together in the desired ratio (e.g., 7:1:1) to achieve maximalcalcium regulation of the actin filament. The screen is conducted at apCa=6.5. This calcium concentration is in the physiological range duringmuscle contraction.

To measure the generation of ADP during the reaction, a pyruvatekinase/lactate dehydrogenase/NADH coupled enzyme system (PK/LDH) isadded to the actin. The myosin is kept separately. The plates are readin real time so that kinetic curves are obtained. These compounds are inDMSO and were already spotted onto the bottoms of 384 well plates at 10to 40 μg/ml final concentration.

Example 3 Actin Preparation

-   -   1. Extract powder (as prepared in Example 6 or 7 below) with 20        ml buffer A (see below, add BME and ATP just prior to use in        each of the following steps) per gram of powder (200 ml per 10        g). Use a large 4 L beaker for 150 g of powder. Mix vigorously        to dissolve powder. Stir at 4° C. for 30 min.    -   2. Separate extract from the hydrated powder by squeezing        through several layers of cheesecloth. Cheesecloth should be        pre-sterilized by microwaving damp for 1-2 min.    -   3. Re-extract the residue with the same volume of buffer A and        combine extracts.    -   4. Spin in JLA10 rotor(s) for 1 hr at 10K rpm (4° C.). Collect        supernatant through 2 layers of cheesecloth.    -   5. Add ATP to 0.2 mM and MgCl₂ to 50 mM. Stir on stir plate at        4° C. for 60 minutes to allow actin to polymerize/form        para-crystals.    -   6. Slowly add solid KCl to 0.6 M (45 g/l). Stir at 4° C. for 30        min.    -   7. Spin in JLA10 rotor(s) at 10K rpm for 1 hr.    -   8. Depolymerization: Quickly rinse surface of pellets with        buffer A and dispose of wash. Soften the pellets by        pre-incubation on ice with small amount of buffer A in each tube        (use less than half of final resuspension volume total in all        tubes). Resuspend by hand first with cell scraper and combine        pellets. Wash tubes with extra buffer using a 25 ml pipette and        motorized pipettor, aggressively removing actin from sides of        tubes. Homogenize in large dounce in cold buffer A on ice. Use 3        ml per gram of powder originally extracted.    -   9. Dialyze against buffer A with 4 changes over 48 hour period.    -   10. Collect dialyzed actin and spin in the 45Ti rotor at 40K rpm        for 1.5 hr (4° C.).    -   11. Collect supernatant (G-Actin). Save a sample for gel        analysis and determination of protein concentration.

To polymerize G-actin for storage add KCl to 50 mM (from 3 M stock),MgCl₂ to 1 mM, and NaN₃ to 0.02% (from 10% stock). Store at 4° C. Do notfreeze.

Buffer A: 2 mM tris/HCl, 0.2 mM CaCl₂, 0.5 mM (36 μl/L)2-mercaptoethanol, 0.2 mM Na₂ ATP (added fresh), and 0.005% N-azide; pH8.0.

Example 4 Powder Preparation

-   -   1. Volumes are given per about 1000 g of the minced muscle.    -   2. Pre-cut and boil cheesecloth for 10 min in water. Drain and        dry.    -   3. Mince chicken breast in a prechilled meat grinder.    -   4. Extract with stirring in 2 L of 0.1 M KCl, 0.15 M        K-phosphate, pH 6.5 for 10 min at 4° C. Spin 5000 rpm, 10 min,        4° C. in JLA. Collect the pellet.    -   5. Extract pellets with stirring with 2 L of 0.05 M NaHCO₃ for 5        min. Spin 5000 rpm, 10 min, 4° C. in JLA. Collect the pellet.        Repeat the extraction once more.    -   6. Extract the filtered residue with 2 L of 1 mM EDTA, pH 7.0        for 10 min with stirring.    -   7. Extract with 2 L of H₂O for 5 min with stirring. Spin 10000        rpm, 15 min, 4° C. in JLA. Carefully collect the pellet, part of        which will be loose and gelatinous.    -   8. Extract 5 times with acetone (2 L of acetone for 10 min each        with stirring). Squeeze through cheesecloth gently. All acetone        extractions are performed at room temperature. Acetone should be        prechilled to 4° C.    -   9. Drying: Place the filtered residue spread on a cheesecloth in        a large glass tray and leave in a hood overnight. When the        residue is dry, put in a wide mouth plastic bottle and store at        20° C.

Example 5 Alternate Powder Preparation

Based on Zot & Potter (1981) Prep. Biochem. 11(4) pp. 381-395.

-   -   1. Dissect left ventricles of the cardiac muscle. Remove as much        of the pericardial tissue and fat as possible. Grind in a        prechilled meat grinder. Weigh.    -   2. Prepare 5 volumes of Extract buffer (see below). Be sure the        pH=8.0. Then, homogenize the meat in a blender, 4 times 15 sec        on blend with 15 secs in between. Do this with 1 volume        weight/volume) of buffer taken from the 5 volumes already        prepared. Add the homogenate back to the extract buffer and stir        until well mixed (5 minutes).    -   3. Filter through one layer of cheesecloth in large        polypropylene strainer. Resuspend back into 5 volumes of extract        buffer as above.    -   4. Repeat Step 3 four more times. At the end, do not resuspend        in extraction buffer but proceed to Step 5. The pellets should        be yellow white.    -   5. Resuspend in 3 volumes (according to original weight) of 95%        cold ethanol. Stir for 5 min and squeeze through cheesecloth as        above, repeat two more times.    -   6. Weigh squeezed residue and then resuspend in 3 volumes (new        weight/volume) of cold diethyl ether.    -   7. Repeat Step 6 a total of three times.    -   8. Leave overnight in a single layer on a cheesecloth in a glass        tray.    -   9. When dry, collect the powder, weigh and store in a wide-mouth        jar at 4° C.        EXTRACT BUFFER: 50 mM KCl, 5 mM Tris pH 8.0

Prepare as 50 times concentrate:

For 2 L

250 mM Tris pH 8.0. Tris Base (121.14 g/mol, 60.6 g)

pH to 8.0 with conc. HCl, then add:

2.5 M KCl (74.55 g/mol, 372 g)

Example 6 Purification of Skeletal Muscle Myosin

See, Margossian, S. S, and Lowey, S. (1982) Methods Enzymol. 85, 55-123and Goldmann, W. H. and Geeves, M. A. (1991) Anal. Biochem. 192, 55-58.

Solution A: 0.3 M KCl, 0.15 M potassium phosphate, 0.02 M EDTA, 0.005 MMgCl₂, 0.001 M ATP, pH 6.5.

Solution B: 1 M KCl, 0.025 M EDTA, 0.06 M potassium phosphate, pH 6.5.

Solution C, 0.6 M KCl, 0.025 M potassium phosphate, pH 6.5.

Solution D: 0.6 M KCl, 0.05 M potassium phosphate, pH 6.5.

Solution E: 0.15 M potassium phosphate, 0.01 M EDTA, pH 7.5.

Solution F: 0.04 M KCl, 0.01 M potassium phosphate, 0.001 M DTT, pH 6.5.

Solution G: 3 M KCl, 0.01 M potassium phosphate, pH 6.5.

All procedures are carried out at 4° C.

-   -   1. Obtain ˜1000 g skeletal muscle, such as rabbit skeletal        muscle.    -   2. Grind twice; extract with 2 L solution A for 15 min while        stirring; add 4 L cold H₂O, filter through gauze; dilute with        cold H₂O to ionic strength of 0.04, (about 10-fold); let settle        for 3 h; collect precipitate at 7,000 rpm in GSA rotor for 15        min.    -   3. Disperse pellet in 220 ml solution B; dialyze overnight        against 6 L solution C; slowly add ˜400 ml equal volume cold        distilled H₂O; stir for 30 min; centrifuge at 10,000 rpm for 10        min in GSA rotor.    -   4. Centrifuge supernatant at 19,000 rpm for 1 h.    -   5. Dilute supernatant to ionic strength of 0.04 (˜8-fold); let        myosin settle overnight; collect about 5-6 L fluffy myosin        precipitate by centrifuging at 10,000 rpm for 10 min in GSA        rotor.    -   6. Resuspend pellet in minimal volume of solution G; dialyze        overnight against 2 L solution D; centrifuge at 19,000 rpm for 2        h, in cellulose nitrate tubes; puncture tubes and separate        myosin from fat and insoluble pellet.    -   7. Dilute supernatant to 5-10 mg/ml and dialyze against solution        E extensively, load onto DEAE-sephadex column.    -   8. Preequilibrate with solution E; apply 500-600 g myosin at 30        ml/h; wash with 350 ml solution E; elute with linear gradient of        0-0.5 M KCl in solution E (2×1 liter); collect 10 ml fractions;        pool myosin fractions (>0.1 M KCl); concentrate by overnight        dialysis against solution F; centrifuge at 25,000 rpm for 30        min; store as above.    -   9. The myosin is then cut with chymotrypsin or papain in the        presence of EDTA to generate the S1 fragment which is soluble at        the low salt conditions optimal for ATPase activity (Margossian        supra).

Example 7

Using procedures similar to those described herein, the compounds in thefollowing table were synthesized and tested.

AC1.4 M/Z (M + 1 unless Name Median denoted)5-bromo-3-(ethylpropyl)imidazo[5,4-b]pyridin-2-ol 8.050 284.0; 286.03-(ethylpropyl)-5-methylimidazo[5,4-b]pyridin-2-ol 43.2993-(ethylpropyl)-5-(methylethyl)imidazo[5,4-b]pyridin- 36.470 248.2 2-ol3-(ethylpropyl)-5-vinylimidazo[5,4-b]pyridin-2-ol 3.836 230.0 (M − H)5-chloro-3-(ethylpropyl)imidazo[5,4-b]pyridin-2-ol 21.134 240.05-(dimethylamino)-3-(ethylpropyl)imidazo[5,4- 2.611 249.2 b]pyridin-2-ol3-(ethylpropyl)-5-ethynylimidazo[5,4-b]pyridin-2-ol 2.366 230.05-((1E)prop-1-enyl)-3-(ethylpropyl)imidazo[5,4- 11.911 246.1b]pyridin-2-ol 3-(ethylpropyl)-5-(1-methylvinyl)imidazo[5,4- 1.413 246.2b]pyridin-2-ol 5-((1Z)prop-1-enyl)-3-(ethylpropyl)imidazo[5,4- 0.840246.1 b]pyridin-2-ol

While some embodiments have been shown and described, variousmodifications and substitutions may be made thereto without departingfrom the spirit and scope of the invention. For example, for claimconstruction purposes, it is not intended that the claims set forthhereinafter be construed in any way narrower than the literal languagethereof, and it is thus not intended that exemplary embodiments from thespecification be read into the claims. Accordingly, it is to beunderstood that the present invention has been described by way ofillustration and not limitations on the scope of the claims.

1. At least one chemical entity selected from compounds of Formula I:

and pharmaceutically acceptable salts and tautomers thereof, wherein R³is selected from optionally substituted alkyl, optionally substitutedcycloalkyl, optionally substituted aryl, optionally substitutedheteroaryl, and optionally substituted heterocycloalkyl; R⁵ is selectedfrom optionally substituted alkenyl and optionally substituted alkynyl;R⁶ is selected from hydrogen, halo, hydroxy, lower alkyl, and lowerhaloalkyl; and R⁷ is selected from hydrogen, optionally substitutedacyl, optionally substituted alkyl, optionally substituted aryl,optionally substituted cycloalkyl, optionally substituted heteroaryl,optionally substituted aminocarbonyl, sulfonyl, sulfanyl, sulfinyl,carboxy, optionally substituted alkoxycarbonyl, hydroxy, and cyano. 2.At least one chemical entity of claim 1 wherein R⁷ is selected fromhydrogen and lower alkyl.
 3. At least one chemical entity of claim 2wherein R⁷ is hydrogen.
 4. At least one chemical entity of claim 1wherein R⁶ is hydrogen.
 5. At least one chemical entity of claim 1wherein R⁵ is selected from alkenyl and alkynyl.
 6. At least onechemical entity of claim 5 wherein R⁵ is selected from prop-1-enyl,ethynyl, vinyl, and 1-methylvinyl.
 7. At least one chemical entity ofclaim 6 wherein R⁵ is selected from ethynyl, (1Z)-prop-1-enyl,(1E)-prop-1-enyl and 1-methylvinyl.
 8. At least one chemical entity ofclaim 1 wherein R³ is selected from optionally substituted cycloalkyland optionally substituted lower alkyl.
 9. At least one chemical entityof claim 8 wherein R³ is lower alkyl optionally substituted with one ormore groups selected from optionally substituted phenyl, hydroxy,optionally substituted alkoxy, optionally substituted amino andoptionally substituted heterocycloalkyl.
 10. At least one chemicalentity of claim 9 wherein R³ is lower alkyl optionally substituted withone or more groups selected from hydroxy, optionally substituted alkoxy,and optionally substituted amino.
 11. At least one chemical entity ofclaim 10 wherein R³ is selected from lower alkyl and lower alkylsubstituted with hydroxyl.
 12. At least one chemical entity of claim 11wherein R³ is pentyl.
 13. At least one chemical entity of claim 1wherein the compound of Formula I is chosen from5-((1Z)prop-1-enyl)-3-(ethylpropyl)imidazo[5,4-b]pyridin-2-ol;5-((1E)prop-1-enyl)-3-(ethylpropyl)imidazo[5,4-b]pyridin-2-ol;3-(ethylpropyl)-5-ethynylimidazo[5,4-b]pyridin-2-ol;3-(ethylpropyl)-5-vinylimidazo[5,4-b]pyridin-2-ol; and3-(ethylpropyl)-5-(1-methylvinyl)imidazo[5,4-b]pyridin-2-ol; or apharmaceutically acceptable salt or tautomer thereof.
 14. Apharmaceutically acceptable composition comprising a pharmaceuticallyacceptable carrier and at least one chemical entity of claim 1 or apharmaceutically acceptable salt or tautomer thereof.
 15. Thepharmaceutical composition of claim 14, wherein the composition isformulated in a form chosen from a tablet, capsule, powder, liquid,suspension, suppository and aerosol.
 16. At least one chemical entity ofclaim 1, wherein R³ is selected from 3-pentyl, 2-methylpropyl,2,2-dimethylpropyl and 2-ethylbutyl.
 17. At least one chemical entity ofclaim 16, wherein R³ is 3-pentyl.
 18. At least one chemical entity ofclaim 16, wherein R⁵ is selected from prop-1-enyl, ethynyl, vinyl and1-methylvinyl.
 19. At least one chemical entity of claim 18, wherein R⁶is hydrogen and R⁷ is hydrogen.